CAJ | 학술논문

目的:在原核系统中高效表达抗TNF-α单克隆抗体VH/K21,以便进一步研究其生物学功能,预测临床应用前景.方法:以已筛选获得的TNF-α抗体基因VH/K21为模板进行PCR扩增,产物与其表达质粒进行双酶切,连接后克隆进入PET22b(+)/BL21(DE3)感受态中并在该原核系统中实现可溶性表达;对表达产物进行Western blotting、ELISA鉴定.结果:成功克隆与表达出可溶性的高效表达的人源TNF-α单克隆抗体.序列分析表明该克隆具有全新的人源TNF-α抗体基因,它可以特异识别人TNF-α.结论:人源TNF-α单克隆抗体的获得为进一步研制应用于临床的TNF-α人源抗体奠定了基础,也将为其他人源抗体的制备提供理论依据和技术基础.
목적:재원핵계통중고효표체항TNF-α단극륭항체VH/K21,이편진일보연구기생물학공능,예측림상응용전경.방법:이이사선획득적TNF-α항체기인VH/K21위모판진행PCR확증,산물여기표체질립진행쌍매절,련접후극륭진입PET22b(+)/BL21(DE3)감수태중병재해원핵계통중실현가용성표체;대표체산물진행Western blotting、ELISA감정.결과:성공극륭여표체출가용성적고효표체적인원TNF-α단극륭항체.서렬분석표명해극륭구유전신적인원TNF-α항체기인,타가이특이식별인TNF-α.결론:인원TNF-α단극륭항체적획득위진일보연제응용우림상적TNF-α인원항체전정료기출,야장위기타인원항체적제비제공이론의거화기술기출.
Objective: To express the TNF-α moneclonal antibody VH/K21 in the prokaryotic expression system, in order to further explore its biological functions and predict its clinical application prospect. Methods: The screened TNF-α an-tibedy VH/K21 gene was amplified by PCR and cloned into the prokaryotie expression vector pET-22b(+)/BL21(DE3), and achieved soluble expression. The products were identified by Western blotting and ELISA. Results: The soluble and high expressed human TNF-α monoclonal antibody was successfully cloned and expressed. Sequence analysis suggested that this clone had fully new human TNF-α antibody gene, and it could specifically recognize human TNF-α. Conclusion: The obtain of human TNF-α monoclonal antibody lays a foundation for its clinical application, and would provide a theoretical and technical basis for the preparation of other human antibodies.